Skip to main content
Log in

Expression of metabolic enzyme genes and heat-shock protein genes during embryonic development in diapause and non-diapause egg of multivoltine silkworm Bombyx mori

  • Published:
Biologia Aims and scope Submit manuscript

Abstract

The expression of metabolic enzyme genes and heat-shock protein genes (Hsp) during early embryogenesis in diapause and non-diapause eggs of the silkworm Bombyx mori was quantified by semi-quantitative RT-PCR. The trehalase gene (Tre) was expressed in non-diapause eggs up-to nine days, while in diapause eggs was not up regulated. The glycogen phosphorylase gene (GPase) was expressed in non-diapause eggs, whereas in diapause eggs a high level was observed in early stage, but down regulated in later stage. The phosphofructokinase gene (PFK) and sorbitol dehyrogenase-2 gene (SDH-2) expression was fluctuated in non-diapause eggs, whereas in diapause eggs these were expressed only at early stage and not observed in later stage. The glucose-6-phosphate dehydrogenase gene (G6P-DH) in non-diapause eggs was highly expressed during the differentiation phase and decreased in the organogenesis phase. In contrast to this, expression in diapause eggs was of low level during differentiation phase and of high level observed in the organogenesis phase. In the tissues, PFK and SDH-2 were selectively expressed in cuticle and midgut, whereas Tre expression was high in midgut and ovary of larvae incubated at 15°C. The Hsp (20.4, 20.8, 40, 70, and 90) were expressed in both diapause and non-diapause eggs. Their expression was, however, selective in tissues with Hsp20.4 in midgut and ovary, Hsp40 in head, Hsp70 in cuticle and Hsp90 in ovary and head in high amounts at 15°C. These results suggest that the metabolic enzyme genes studied except Hsp play a major role during embryogenesis of diapause and non-diapause silkworm.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

DH:

diapause hormone

GPase:

glycogen phosphorylase

G6P-DH:

glucose-6-phosphate dehydrogenase

Hsp:

heat-shock protein

PFK:

phosphofructokinase

SDH:

sorbitol dehydrogenase

Tre:

trehalase

References

  • Andrewartha H.G. 1952. Diapause in relation to the ecology of insects. Biol. Rev. 27: 50–107.

    Article  Google Scholar 

  • Benson D.A., Karsch-Mizrachi I., Lipman D.J., Ostell J. & Wheeler D.L. 2007. GenBank. Nucleic Acids Res. 35 (Data-base Issue): D21–D25.

    Article  PubMed  CAS  Google Scholar 

  • Chen C.P. & Denlinger D.L. 1990. Activation of phosphorylase in response to cold and heat stress in the flesh fly Sarcophaga crassipalpis. J. Insect Physiol. 36: 549–553.

    Article  CAS  Google Scholar 

  • Coulon B.M. & Mathelin J. 1991. Variations in the rates of synthesis of heat shock proteins, Hsp70, between laying and neurula in the diapausing embryo of the silkworm, Bombyx mori. Sericologia 31: 295–300.

    Google Scholar 

  • Coulon M. & Dorel C. 1991. The arrest of embryogenesis at gastrula stage in the diapausing silkworm Bombyx mori is related to the synthesis of protein p61. Dev. Genes Evol. 199: 469–475.

    Google Scholar 

  • Denlinger D.L., Rinehart J.P. & Yocum G.D. 2001. Stress proteins: a role in insect diapause?, pp. 155–171. In: Denlinger D.L., Giebultowicz J.M. & Saunders D.S. (eds), Insect Timing: Circadian Rhythmicity to Seasonality, Elsevier, Amsterdam.

    Chapter  Google Scholar 

  • Feder M.E. & Hofmann G.E. 1999. Heat shock proteins, molecular chaperones, and the stress response: evolutionary and ecological physiology. Annu. Rev. Physiol. 61: 243–282.

    Article  PubMed  CAS  Google Scholar 

  • Fujiwara Y., Shindome C., Takeda M. & Shiomi K. 2006. The role of ERK and p38 MAPK signaling cascades on embryonic diapause initiation and termination of the silkworm Bombyx mori. Insect Biochem. Mol. Biol. 36: 47–53.

    Article  PubMed  CAS  Google Scholar 

  • Furusawa T. & Yang W.J. 1987. Fluctuation of the free sugar with embryonic development of non-diapause eggs of the silkworm, Bombyx mori. J. Sericult. Sci. Japan 56: 143–149.

    CAS  Google Scholar 

  • Goto S.G. & Kimura M.T. 2004. Heat-shock-responsive genes are not involved in the adult diapause of Drosophila triauraria, Gene 326: 117–122.

    Article  PubMed  CAS  Google Scholar 

  • Hayakawa Y. & Chino H. 1982. Phosphofructokinase as a possible key enzyme regulating glycerol or trehalose accumulation in diapausing insects. Insect Biochem. 12: 639–642.

    Article  CAS  Google Scholar 

  • Hayward S.A.L., Rinehart J.P. & Denlinger D.L. 2004. Desiccation and rehydration elicit distinct heat shock protein transcript responses in flesh fly pupae. J. Exp. Biol. 207: 963–971.

    Article  PubMed  CAS  Google Scholar 

  • Horie Y., Kanda T. & Mochida Y. 2000. Sorbitol as an arrester of embryonic development in diapausing eggs of the silkworm, Bombyx mori. J. Insect Physiol. 46: 1009–1016.

    Article  PubMed  CAS  Google Scholar 

  • Hwang J.S., Go H.J., Goo T.W.S., Yun E.Y., Ahn M.Y., Kim S.R., Park K.H., Kim I., Jeon J.P. & Kang S.W. 2007. Molecular characterization of small heat shock protein (Hsp 20.8A) from the silkworm, Bombyx mori. Int. J. Indust. Entomol. 15: 75–78.

    Google Scholar 

  • Hwang J.S., Go H.J., Goo T.W., Yun E.Y., Choi K.H., Su I.S., Lee S.M., Lee B.H., Kim I., Chun T. & Kang S.W. 2005. The analysis of differentially expressed novel transcripts in diapausing and diapause-activated eggs of Bombyx mori. Arch. Insect Biochem. Physiol. 59: 197–201.

    Article  PubMed  CAS  Google Scholar 

  • Joplin K.H., Yocum G.D. & Denlinger D.L. 1990. Cold shock elicits expression of the heat shock protein in the flesh fly Sarcophaga crassipalpis. J. Insect Physiol. 36: 825–834.

    Article  CAS  Google Scholar 

  • Kitazawa T., Kanda T. & Takami T. 1963. Changes of mitotic activity in silkworm eggs in relation to diapause. Bull. Sericult. 18: 283–285.

    Google Scholar 

  • Kostal V., Tollarova M. & Sula J. 2004. Adjustments of the enzymatic complement for polyol biosynthesis and accumulation in diapausing cold-acclimated adults of Pyrrhocoris apterus. J. Insect Physiol. 50: 303–313.

    Article  PubMed  CAS  Google Scholar 

  • Krishnaswami S. 1978. New Technology of Silkworm Rearing. Bulletin No. 2. Central Sericultural Research and Training Institute, Mysore, Central Silk Board, Government of India.

  • Lindquist S. & Craig E.A. 1988. The heat shock proteins. Annu. Rev. Genet. 22: 631–677.

    Article  PubMed  CAS  Google Scholar 

  • Moribe Y., Niimi T., Yamashita O. & Yaginuma T. 2001. Samui, a novel cold inducible gene, encoding a protein with a BAG domain similar to silencer of death domains (SODD/BAG-4) isolated from Bombyx diapause eggs. Eur. J. Biochem. 268: 3432–3442.

    Article  PubMed  CAS  Google Scholar 

  • Niimi T., Yamashita O. & Yaginuma T. 1993. A cold-inducible Bombyx gene encoding a protein similar to mammalian sorbitol dehydrogenase: yolk nuclei-dependent gene expression in diapause eggs. Eur. J. Biochem. 213: 1125–1131.

    Article  PubMed  CAS  Google Scholar 

  • Rinehart J.P. & Denlinger D.L. 2000. Heat shock protein 90 is down regulated during pupal diapause in the flesh fly, Sarcophaga crassipalpis, but remains responsible to thermal stress. Insect Mol. Biol. 9: 641–645.

    Article  PubMed  CAS  Google Scholar 

  • Rinehart J.P., Li A., Yocum G.D., Robich R.M., Hayward A.L. & Denlinger D.L. 2007. Up regulation of heat shock protein is essential for cold survival during insect diapause. Proc. Natl. Acad. Sci. USA 104: 11130–11137.

    Article  PubMed  CAS  Google Scholar 

  • Rinehart J.P., Yocum G.D. & Denlinger D.L. 2000. Developmental up regulation of inducible Hsp 70 transcripts, but not the cognate form, during pupal diapause in the flesh fly, Sarcophaga carssipalsi. Insect Biochem. Mol. Biol. 30: 515–521.

    Article  PubMed  CAS  Google Scholar 

  • Sawada H., Yamahama Y., Yamamoto T., Mase K., Ogawa H. & Ino T. 2006. A novel RNA helicase like protein during early embryonic development in silkworm Bombyx mori; Molecular characterization and intracellular localization. Insect Biochem. Mol. Biol. 36: 911–920.

    Article  PubMed  CAS  Google Scholar 

  • Sonoda S., Fukumoto K., Izumi Y., Ashfaq M., Yoshida H. & Tsumaki H.A. 2006. Small Hsp gene is not responsible for diapause and cold tolerance acquisition in Chilo suppressalis. J. Appl. Entomol. 130: 309–313.

    Article  CAS  Google Scholar 

  • Storey K.B. & Storey J.M. 1988. Freeze tolerance in animals. Physiol. Rev. 68: 27–84.

    PubMed  CAS  Google Scholar 

  • Storey K.B. & Storey J.M. 1991. Biochemistry of cryoprotectants, pp. 64–93. In: Denlinger D. & Lee R.E. (eds), Insects at Low Temperature, Chapman and Hall, New York.

    Google Scholar 

  • Su Z.H., Ikeda M., Sato Y., Imai K., Isobe M. & Yamashita O. 1994. Molecular characterization of ovary trehalase of the silkworm, Bombyx mori and its transcriptional activation by diapause hormone. Biochim. Biophys. Acta 1218: 366–374.

    PubMed  CAS  Google Scholar 

  • Su Z.H., Sato Y. & Yamashita O. 1993. Purification, cDNA cloning and Northern blot analysis of trehalase of pupal midgut of the silkworm, Bombyx mori. Biochim. Biophys. Acta 1173: 217–224.

    PubMed  CAS  Google Scholar 

  • Suzuki M.G., Terada T., Kobayashi M. & Shimada T. 1999. Diapause-associated transcription of BmEts, a gene encoding an ETS transcription factor homolog in Bombyx mori. Insect Biochem. Mol. Biol. 29: 339–347.

    Article  PubMed  CAS  Google Scholar 

  • Trang L.T.D., Sehadova H., Ichihara N., Iwai S., Mita K. & Takeda M. 2006. Casein kinases I of the silkworm, Bombyx mori: their possible roles in circadian timing and developmental determination. J. Biol. Rhythms 21: 335–349.

    Article  CAS  Google Scholar 

  • Tsimuki H., Rojas R.R., Storey K.B., Baust J.G. 1987. The fate of (14C) glucose during cold-hardening in Eurosta solidiaginis (Fitch). Insect Biochem. 17: 347–352.

    Article  Google Scholar 

  • Wood F.B. & Nordin J.H. 1980. Activation of the hexose monophosphate shunt during cold induced glycerol accumulation by Protophormia terranoval. Insect Biochem. 10: 87–93.

    Article  CAS  Google Scholar 

  • Yaginuma T., Kobayashi M. & Yamashita O. 1990. Distinct effects of different low temperatures on the induction of NAD sorbitol dehydrogenase activity in diapause eggs of the silkworm, Bombyx mori. J. Comp. Physiol. 160: 277–285.

    CAS  Google Scholar 

  • Yaginuma T. & Yamashita O. 1978. Polyol metabolism related to diapause in Bombyx eggs: different behaviour of sorbitol from glycerol during diapause and post-diapause. J. Insect Physiol. 24: 347–354.

    Article  CAS  Google Scholar 

  • Yaginuma T. & Yamashita O. 1979. NAD-sorbitol dehydrogenase activity in relation to the termination of diapause in eggs of Bombyx mori. Insect Biochem. 9: 547–553.

    Article  CAS  Google Scholar 

  • Yaginuma T. & Yamashita O. 1999. Oxygen consumption in relation to sorbitol utilization at the termination of diapause in eggs of the silkworm Bombyx mori. J. Insect Physiol. 44: 621–627.

    Article  Google Scholar 

  • Yamamoto T., Kanekatsu M., Nakagoshi M., Kato T., Mase K. & Sawada H. 2005. Casein kinase 2 during early embryonic development in silkworm Bombyx mori: cDNA sequence, gene expression, and enzyme activity. DNA Seq. 16: 446–455.

    PubMed  CAS  Google Scholar 

  • Yamashita O. 1996. Diapause hormone of the silkworm, Bombyx mori: structure, gene expression and function. J. Insect Physiol. 42: 669–679.

    Article  CAS  Google Scholar 

  • Yamashita O. & Hasegawa K. 1985. Embryonic diapause, pp. 407–434. In Kerkut G.A. & Gilbert L.I. (eds), Comprehensive Insect Physiology, Biochemistry and Pharmacology, Pergamon Press, Oxford.

    Google Scholar 

  • Yamashita O., Suzuki K. & Hasegawa K. 1975. Glycogen phosphorylase activity in relation to diapause initiation in Bombyx egg. Insect Biochem. 5: 707–718.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Kangayam M. Ponnuvel.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Saravanakumar, R., Ponnuvel, K.M. & Qadri, S.M.H. Expression of metabolic enzyme genes and heat-shock protein genes during embryonic development in diapause and non-diapause egg of multivoltine silkworm Bombyx mori . Biologia 63, 737–744 (2008). https://doi.org/10.2478/s11756-008-0124-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2478/s11756-008-0124-x

Key words

Navigation